The Genus Ferulago: A Review on Ethnopharmacology, Phytochemistry, and Pharmacology

The Ferulago genus appertains to the Umbelliferae family comprises 49 species which are mainly distributed in Asia, Europe, and Africa. This paper aims to review the morphological properties of Ferulago species, herbal components, and their pharmacological properties. The information of this review paper has been collected from journals available in databases including Science Direct, Web of Science, Scopus, PubMed, EBSCO, Google Scholar, and Hindawi up to March 2020. In traditional medicine, the genus of Ferulago has been used to treat intestinal worms, snake bites, wound skin infections, diseases of the spleen and gastrointestinal tract, and headaches. It not only has been used traditionally as a preservative agent to dairy, oil ghee, and meat but also has given them a pleasant taste. The main components of Ferulago spp. are monoterpenes, sesquiterpenes, coumarin, furanocoumarin, flavonoids, and terpenoids have been the reason for the antimicrobial, antioxidant, anticoagulant, antidiabetic, Alzheimer, and larvicidal properties of this plant. This review confirms that many traditional uses of some Ferulago species have now been validated by modern pharmacology research. Rigorous investigations of all the species of Ferulago concerning phytochemical and pharmacological properties, mainly their mechanism of action, efficacy, and safety might offer a context for researchers to prosper plant-derived medications like anti-diabetes, antibiotics, and sedatives treating drugs, and the key to directing clinical trials.


Introduction
Herbal used in traditional and modern medicine presents a valuable source of secondary metabolites and different pharmacological and biological activities. Hence, they could be utilized as a lead compound to produce new drugs and treat numerous diseases (1). The Apiaceae or Umbelliferae, commonly known as the parsley, carrot, or celery family, is one of the biggest plant families in the world usually characterized by aromatic plants with hollow stems (2). Several plants belonging

The botany of Ferulago plants
The Ferulago genus appertains to the Umbelliferae family and comprises 49 species which are mainly distributed in Asia, Europe, and Africa (8). The genus Ferulago is distributed in Turkey with 34 species, out of which 18 are endemic (9). The 8 species exist in Iran, out of which three are endemic (10). According to The Plant List website, the genus Ferulago consists of 48 accepted species names and 13 unassessed species (11). Table 1 depicts the name of accepted species and their synonyms. Ferulago genus is 60-150 cm tall; annual or perennial plants grow at altitudes of 1900-3200m above sea level with yellow fruits. They are characterized by persistent bracts and bracteoles and small flowers which are widely distributed in Turkey, Iraq, and west of Iran (from the flora of Iran, particularly in Kermanshah, Ilam, Lorestan, and Kurdistan) (8,12,13). Pictures of Ferulago bernardii and fruits of ferulago stellata are shown in Fig.1. The high distribution of Ferulago spp. in these areas suggests that the core center of the biodiversity of the genus Ferulago is in Anatolia. Based on taxonomy the species of this genus resemble Prangos and Ferula species. They are widely used in the Anatolia region for several purposes (7,14). The most studied species is F. angulata which is divided based on ovaries, the flowering of in fluorescence, and fibers (rather absence or presence of trichomes) into two subspecies: F. angulate subsp. angulata and F. angulate subsp. Carduchorum by Chamberlain in 1987 (15).

Traditional use of Ferulago spp.
Ferulago like Ferula and Prangos species have been traditionally utilized back in history in folk medicine for the medicament of hemorrhoids, and intestinal worms, peptic, sedative, carminative, digestive, aphrodisiac, snakebite, wound skin infections, headache, diseases of the gastrointestinal tract and spleen (5,16). Due to the aromatic property of Ferulago, some of them have been applied in Iranian tradition as spices in different foods, such as dairy and meat products, for increasing the flavor and as an aroma. They have also been employed as natural preservatives for enhancing food expiration dates (8, 16 and 17). Bakhtiari nomads use F. angulata to make some foods and Nomads of Fars province utilize this plant to flavor yogurt. The F. angulata is also used as a sedative, food-digestive, tonic, antibacterial, and antiparasitic (18). In Turkish traditional medicine, the aerial parts of a few members of this genus are used as an immunostimulant, tonic, sedative, digestive, anti-bronchitis, flavor, vermicidal, anthelmintic, and antipeptic (19,20). Likewise, the root parts are used for the treatment of dermatological 354 The genus Ferulago, a Review

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The genus Ferulago, a Review disorders and cancers and as an aphrodisiac and are preferred as fodder to improve animal productivity (7). The seeds are applied for eye pains in the form of inhalation (8). It has been reported that dried or fresh leaves of Ferulago are used as foot deodorant by indigenous people of the north of Iraq (21).

Phytochemistry of essential oils from the genus Ferulago species
Essential oils or volatile oils are the phytochemical complexes of different aromatic components, mainly monoterpenoids and sesquiterpenoids, which are obtained from plant materials, for instance, leaves, fruits, seeds buds, flowers, roots, and bark. They are characterized by having a strong odor generally lower density than water, being volatile, rarely colored, liquid, lipophilic, and soluble in organic solvents (7). Despite the way that these volatile oils involve around 20-60 constituents, just a few of them exist at high amounts (20-70%) in correlation with different constituents existing in low sums (22). These phytochemicals assume a vital part in the protection of the herbs from herbivores, insects, bacteria, fungi, viruses, and also help to attract pollinators (23). These essential oils could be extracted with conventional methods (steam distillation, hydrodistillation (HD), organic solvent extraction) and innovative techniques (In situ microwave-generated hydrodistillation, supercritical carbon dioxide, microwave steam diffusion, microwave hydro diffusion and gravity, and microwave steam distillation). Among these methods, the hydrodistillation method is the most prevalent technique for obtaining essential oils (7, 24 and 25). The method of extraction, drying methods, genotypic variation, geographical origin of the plant, stage of the development, and part of the plant used may drastically affect the composition of essential oils of plants (26)(27)(28)     main compounds of these essential oils, part of the herb that was used, methods of extraction, percentage yield, class of components, and their references. According to data of the volatile oils extracted from various Ferulago spp., it is clear that every species and each part of the plant have a diversified set of main compounds. Therefore, it is hard to find the similarity among the species of this genus concerning the chemicals. Several compounds, like α-Pinene, p-Cymene 2,3,6-trimethyl benzaldehyde, cis-Chrysanthenyl acetate, α-Phellandrene, Sabinene, (Z)-β-Ocimene, Limonene, Myrcene, Terpinolene, Nonacosane, and δ-Cadinene, have been detected as main components of the volatile oils of many Ferulago species. To date, analysis has shown that the α-Pinene is a major compound of several Ferulago species; hence, this might be regarded as a perpetual constituent for the genus (7). There are certain other notable points to report; primarily, the compound of spathulenol was the major component from aerial parts of only two species namely, F. sylvatica and F. thyrsiflora (45). Secondly, the compound of ferulagone was found as the main compound of only F. thirkeana obtained from its fruits (59). Thirdly, carvacrol methyl ether obtained with MD from fruits of F. macrosciadia and F. idaea was the major component of only these two species (47). Furthermore, Cecchini et al. compared the composition of the volatile oil from roots and fruits of F. campestris from two sites of collection and two periods of time. They found significant differences in the percentages of (2, 30 and 6) -trimethyl benzaldehyde (14.8-27.9% in the roots gathered in the summer, 65.2% in roots gathered in the fall) and α-Pinene (58.3-75% in the roots gathered in the summer, 19.3% in the roots gathered collected in the fall) (23).

Phytochemistry of plant extracts from the genus Ferulago species
Phytochemical investigations on Ferulago species have shown the presence of various secondary metabolites including coumarins, coumarin esters, furanocoumarins, aromatic compounds, monoterpenes, sesquiterpenes, flavonoids, quinones, and stilbene. Table  3 depicts the main phytochemicals that have been isolated and characterized from Ferulago species. Up to now, several studies have been done to distinguish active compounds from different parts of the ferulago genus, from which about 73 (three simple coumarins, sixteen furanocoumarins, five dihydro-furanocoumarin, four sesquiterpene coumarin, twelve prenylated coumarins, six pyranocoumarin, nine flavonoids, and eighteen miscellaneous compounds) bioactive compounds were isolated. Based on the literature, coumarins and their derivatives are the most prevalent secondary metabolites on the Ferulago species and might be used as a chemotaxonomic marker in the genus Ferulago (65). The classification of different types of coumarins and various biological applications of each compound was reviewed by Venugopala et. al. (66).

Pharmacological activities
In the last two decades, many ferulago species have been widely studied with advanced scientific methods and reported for several pharmacological properties such as antibacterial activity, antioxidant activity, antidiabetic activity, larvicidal activity, Alzheimer, and anticancer activity. These pharmacological activities of Ferulago are considered to be attributed mainly to its coumarins and furanocoumarins and essential oil (66). Table 4 depicts pharmacological activity and model of study of the genus Ferulago.

Antibacterial activity
The volatile oils of many herbs of genus Ferulago have been the focus on pharmacological activity, particularly from an anti-oxidant, antimicrobial and antifungal point of view (7, 20 and 29). In the literature, antimicrobial and antifungal properties of essential oil were screened versus Gram-positive (Staphylococcus aureus, S. epidermidis, and methicillin-resistant S. aureus), and Gram-negative (Escherichia coli, Salmonella typhimurium, Bacillus cereus, Proteus vulgaris, Enterobacter aerogenes, and Pseudomonas aeruginosa) bacteria, and the yeast (Candida albicans, C. parapsilosis, and C. tropicalis) via broth microdilution assay. Sucu et al., 2019, studied the antimicrobial effects of volatile oil from the roots and aerial parts of F. sandrasica and found that both essential oils were not active against C. tropicalis and C. parapsilosis compared to positive controls (55 (82)(83)(84)(85) Continued  (82,83) Continued fraction of roots, and methanol extracts of roots and aerial parts from F. trachycarpa (62.5 µg/mL) and prantschimgin (31.25 µg/ mL) (78). According to them, the E. coli was less affected than the other microorganisms; the best activity against C. albicans (MIC = 31.25 μg/mL) was obtained by the CH 2 Cl 2 fraction of aerial portions from F. pachyloba and isolated compound prantschimgin. Furthermore, Pinto et al. evaluated the antifungal effects of an essential oil, and two main compounds of it on germ tube formation, ergosterol biosynthesis, and mitochondrial function. Limonene presented a weaker activity (0.32 to 20 μL/mL) than the essential oil and α-pinene with low and similar to MIC and MFC values against the tested organisms (0.08 to 5.0 μL/mL). The essential oil of F. capillaris suppressed germ tube formation at sub-inhibitory dose on Candida albicans. The mechanism of antifungal activity of F. capillaris indicated no distribution on the ergosterol content and defect of mitochondrial role in a dose-dependent way in essential oiltreated C. albicans (62).

Anti-oxidant effect
In recent decades, extensive studies have been conducted on the evaluation of the antioxidant function of medicinal plants as a source of natural compounds not only to combat several degenerative disorders, including cardiovascular disease and cancer, but also as a substitute compound to artificial additives, like butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) in food productions (94). Anti-oxidant constituents diminish the extent of oxidative damage via acting as free radical scavengers (95). There are numerous methods, such as radical scavenging power, reducing power, and inhibition of lipid peroxidation in a β-carotenelinoleate system for assessing the antioxidant activity of plant extracts or volatile oils (96). The DPPH radical-scavenging method is one of the easiest and rapid tests for evaluating the antioxidant activity of natural compounds (16). According to literature, an antioxidant activity study has been performed on F.macrocarpa, F. carduchorum (97), F. bernardii (16), F. sandrasica, F.macedonica, F. trifida (64), F. subvelutina (74), F. cassia (8), F. angulata, and F. campestris (79). The antioxidant activities of four fractions and crude extract of aerial portions of F. carduchorum at 2 vegetative periods (flower and fruit) were assessed using the DPPH method. The best activity belonged to flower crude extract (IC 50 = 0.44 mg/mL) (97). Shahbazi and Shavisi investigated the antioxidant activities of nonpolar and polar sub-fractions of methanolic extract, and the

Alzheimer's disease (AD)
Alzheimer's is one of the neurological disorders with decreasing acetylcholine followed by the deterioration of short-term memory, which occurs in elderly people mostly based on genetic inheritance. One promising strategy for combating AD is using anticholinesterases or acetylcholinesterase inhibitors (AChEIs) to inhibit the hydrolysis of acetylcholine and raise its level in the synaptic cleft (82). Hajimehdipoor and co-worker studied the acetylcholinesterase inhibitory property of the total extracts and fractions of the aerial portions of F. subvelutina and the whole plant of F. angulata employing the Ellman method. Their result implied that a total extract of both Ferulago genes can inhibit the acetylcholinesterase enzyme with 19.7% and 15.8% respectively for F. subvelutina and F. angulata. The dichloromethane fraction displayed the highest AChEI activity among all the fractions (98). Golfakhrabadi et al. found that all the coumarins obtained from the aerial parts of F. carduchorum have AchE enzyme inhibition, among which xanthotoxin revealed the highest inhibitory (IC 50 = 39.64 µM) (70). The essential oil of F. carduchorum has been also found to be of AChE inhibitory activity (IC 50 = 23.6 µL mL -1 ) (34). In another study on AChE inhibitors via bioassay-guided isolation, the dichloromethane extract from the root of F. campestris was investigated. Three daucane ester derivatives (1-acetyl-5-angeloyl lapiferol, ferutinin, and siol anisate), two phenol derivatives (epielmanticine and 2-epilaserine), one polyacetylene (9-epoxyfalcarindiol), and three coumarin derivatives (coladin, coladonin, umbelliprenin) were isolated. All the obtained constituents could inhibit the AChE, but at higher doses (IC 50 1.2-0.1 mM) than the standard galantamine (6.7 μM) and the most active compounds were the epielmanticine and the siol anisate with IC 50 of 0.172 and 0.175 µM, respectively (82). In addition, Karakaya and co-workers studied the anticholinesterase effects of the fractions and extracts from the aerial parts and roots of F. isaurica and F. syriaca. Strong inhibitory activities against butyrylcholinesterase (BuChE) (88.56 ± 2.34%) and AChE (46.99%) at 20 µg/mL were observed in the CHCl 3 part of the root of F. Isaurica. Felamidin (77.11%); prantschimgin (74.82%), two obtained constituents from the chloroform part of roots, presented a strong inhibitory effect against BuChE (19). In another study, AChE inhibitory activities of the volatile oils of the fruits, roots, and flowers of F. trifida were investigated. The result revealed significant AChE inhibitory activity (78.7, 74.3, and 72.1% inhibition of Ach Enzyme, respectively) (64). Hritcu and coworker found the volatile oil extracted from the aerial part of F. angulata has an anti-amnesic activity in scopolamine-induced memory deterioration in rats and diminish AChE activity in hippocampal (99). In another study the volatile oil from fruits of F.pauciradiata revealed strong inhibitory properties against BuChE and AChE (IC 50 = 0.567, 7.987 L/mL, respectively) (20). Anti-diabetic effects α-glucosidase, and α-amylase inhibitory activity of the extracts and some compounds from the roots of F. blancheana, F. trachycarpa, and F. pachyloba have been evaluated by in-vitro bioassay-guided isolation methods to determine the antidiabetic properties of this plant. The obtained result demonstrated that the highest activities versus α-glucosidase with an IC 50 value of 0.3, 2, 2 mg/mL belonged to CH 2 Cl 2 extracts of the roots of F. trachycarpa, F. pachyloba, and F. blancheana, respectively. Suberosin and felamidin compounds possessed momentous α-glucosidase inhibitory activities with IC 50 values of 0.9 and 0.4 mg/mL, respectively, while the IC 50 for acarbose as standard was 4.9 mg/mL. The acarbose depicted a strong α-amylase inhibitory activity (82.3%) at a dose of 1 mg/mL whereas none of the other extracts displayed a significant α-amylase inhibitory effect (100).

Anti-coagulant activity
Golfakhrabadi et al. evaluated the toxicological profile of oral application of the F. carduchorum extract and the anticoagulant effects of two isolated coumarins (suberenol and suberosin) in male Wistar rats. The LD 50 of the plant extract for acute toxicity was over 2000 mg/kg and there were no substantial variations (p > 0.05) among the control and the treated groups concerning the biochemical and hematological parameters. The prothrombin time (PT) of the treated group with a total extract from the aerial part of the plant has not shown a major impact relative to the control group (receiving tap water by gavage) (p > 0.05) at doses of 250 and 500 mg/kg. Meanwhile, suberosin expanded the PT at doses of 3 and 6 mg/kg (16.7 and 17.4 s, respectively) and suberenol at the same dose (16.5 and 17.1 s, respectively) (17).

Anti-malaria
Khanavi et al. assessed the larvicidal activity of chloroform, methanol, ethyl acetate, and the total 80% methanol extract from the aerial part of F. carduchorum against late 3 rd , and early 4 th instar larvae of the malaria vector Anopheles stephensi. The LC 90 of chloroform, ethyl acetate, the total extract, and methanol fractions were 0.455, 1.892, 1.509, and 10.886 ppm, respectively. Moreover, the LC 50 of chloroform, ethyl acetate, the total extract, and methanol fractions were 0.236, 0.744, 0.480, and 3.702 ppm, respectively. The chloroform fraction indicated lower LC 50 and LC 90 values than the other extracts, which might be due to the presence of high content of phytosteroids and coumarins (101). In another study, the larvicidal activity of a few isolated coumarin, methanol, and chloroform extracts of the roots, leaves, and fruits of F. trifida were investigated on the third instar larvae of A. stephensi Listonas. Strong insecticidal properties were found for methanol extract of the fruit with LC 90 and LC 50 values of 18.12 and 2.94 ppm, respectively. Among pure compounds, oxypeucedanin presented moderate toxicity against A. stephensi with LC 90 and LC 50 values of 346.41 and 116.54 ppm, respectively. It could be concluded that the methanol extract from the fruit of F. trifida might be utilized as an effective bio-insecticide in green control programs of mosquitoes, particularly A. stephensi (102).

Aphrodisiac activity
An in-vitro study was conducted by Ozturk et al. to investigate the aphrodisiac activity of the lyophilized water extract from the roots of F. syriaca on the human corpus cavernosum. For finding the mechanism involved in relaxation, the effect of the extract was investigated on the relaxing responses to selective guanilate cyclase inhibitor (oxadiazolo [4,3-α] quinoxalin-1one (ODQ)), NO-synthesis inhibitor (NG-nitro-L-arginine methyl ester (L-NAME)), forskolin, Electrical field stimulation (EFS), sodium nitroprusside, and acetylcholine. The result displayed that the extract could have to relax in a dosedependent way on corpus cavernosum strips, and the L-NAME (small dosages) and QDQ (in all dosages) were able to suppress the extract-induced relaxation in the human corpus cavernosum. The F. syriaca extract enhanced the relaxation response of strips with Ach incubation while the extract did not affect the 372 The genus Ferulago, a Review relaxation induced by EFS, forskolin sodium, and nitroprusside. The result suggested that the extract of F. syriaca could persumabely act via stimulating the NO-cyclic guanosine monophosphate (cGMP) pathway (103).

Toxicity profile of the genus Ferulago
Nowadays, the usage of herbal remedies has increased around the world, and patients falsely feel that they are healthy because they are natural. While these products comprise several bioactive compounds and might cause adverse effects on consumers (104,105). Therefore, it is essential to herbal medicines undergo current safety and efficacy tests. Based on the literature review, there is only one piece of scientific study about the safety of this herb. Golfakhrabadi et al. investigated oral acute, and sub-chronic toxicities of the total extract of the aerial parts of F. carduchorum in the rat (17). To study acute toxicity five rats were treated with one dose of the total extract (2000 mg/kg) orally and the control group was treated with tap water. For the sub-chronic study, the doses (250, 500, and 1000 mg/kg) of the total extract were applied to treated groups by gavage for thirty following days. During the acute toxicity study, the animals did not show any signs of side effects, mortality, and the LD 50 was over 2000 mg/kg in rats. The result of sub-chronic toxicity demonstrated that the total extract did not yield momentous changes in behavior, water, and food intake, breathing, body weight gain, blood metrics, and gastrointestinal properties in rats (17). This study indicated that F. carduchorum may be a healthy additive for conventional applications.

Conclusion and future perspective
To date, around 73 molecules, including coumarin, furanocoumarin, flavonoids, and stilbene have been isolated from the Ferulago species. Among the isolated constituents, mostly are phenolic compounds in which three simple coumarins, sixteen furanocoumarins, five dihydro-furanocoumarin, four sesquiterpene coumarin, twelve prenylated coumarins, six pyranocoumarin, nine flavonoids, and eighteen miscellaneous compounds. Coumarins and their derivatives are considered to be important taxonomic markers of the genus Ferulago. In the pharmacological effect part, we have discussed various studies to evaluate the ethnomedicinal uses and this has shown that the isolated constituents and different extracts from the Ferulago species possess several pharmacological effects including antibacterial, antioxidant, Alzheimer, antidiabetics, anti-malaria, anti-coagulant, aphrodisiac effects. Accessible information regarding Ferulago spp. allows us to explore their potential benefits, highlight the gaps in our knowledge, and conduct future researches to develop new drugs. It can be concluded that the pharmacological study of Ferulago spp. is in agreement with the ethnomedicinal uses of the plants.